Corn inbreds like far045 and hybrids thereof

ABSTRACT

A maize inbred line having the characteristics selected from those of FAR045, including plant parts, tissue, and pigments of the inbred is disclosed.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119 (e) to, andhereby incorporates by reference, U.S. Provisional Application No.60/945,430, filed Jun. 21, 2007. This application hereby incorporates byreference co-pending and concurrently filed U.S. Application AttorneyDocket No. 612.001US1, entitled ANTHOCYANIN PIBMENT/DYE COMPOSITIONS ANDMETHOD OF PROVIDING COMPOSITION THROUGH EXTRACTION FROM CORN.

FIELD OF THE INVENTION

The present invention relates generally to the production of corn andparticularly to the production of inbred corn lines havingcharacteristically red colored plant parts and tissues exhibiting goodpigment stability. The invention also relates to the use of the inbredsin the production of hybrid corn plants and parts and tissue of thesehybrid corn plants and to methods of extracting these pigments fromthese tissues.

BACKGROUND OF THE INVENTION

Maize or corn (Zea mays L.) is a major annual crop species grown forgrain and forage. A monocot, maize is a member of the grass family(Gramineae) and bears seeds in female inflorescences (usually calledears) and pollen in separate male inflorescences (usually calledtassels).

In the U.S., maize is almost exclusively produced by growing hybridvarieties (cultivars). Maize hybrids are typically produced by seedcompanies and sold to farmers. On farms, maize hybrids are usually grownas a row crop. During the growing season herbicides are widely used tocontrol weeds; fertilizers are used to maximize yields; and fungicidesand insecticides are often used to control disease pathogens and insectpests. Before maturity, maize plants may be chopped and placed instorage where the chopped forage undergoes fermentation to become silagefor livestock feed. At maturity in the fall, the seeds are harvested asgrain. The grain may be directly fed to livestock or transported tostorage facilities. From storage facilities, the grain is transported tobe used in making an extremely large number of products, including foodingredients, pigments, snacks, pharmaceuticals, sweeteners, and paperproducts (see, e.g., S. A. Watson and P. E. Ramstad, Eds., Corn:Chemistry and Technology, American Association of Cereal Chemists, Inc.,St. Paul, Minn. (1987)).

While the agronomic performance of maize hybrids has improved, there isa continuing need to develop better hybrids with increased and moredependable pigment and grain and stover yields. Moreover, heat anddrought stress and continually changing insect pests and diseasepathogens present hazards to farmers as they grow maize hybrids. Thus,there is a continual need for maize hybrids which offer higher grainyields in the presence of heat, drought, pathogens and insects.

In accordance with the present invention and in addition to theforegoing agronomic traits, an important characteristic of inbred linesand hybrid plants obtained using the inbred lines of this inventioninvolves pigmentation, such as anthocyanins, the extraction thereof, anduse.

SUMMARY OF THE INVENTION

By means of the present invention, there is provided inbred lines ofcorn having plant parts and tissues with high concentrations of a stablered coloration, which is uncharacteristic of known corn plants usedcommercially today. These desirable attributes are characterized byinbred corn lines such as FAR601 and FAR045.

One aspect of the invention involves the plant parts and tissue of theseinbreds and the use of the plant parts and tissue of these inbred in theextraction and use of the red plant pigments associated with theseinbreds and their hybrids.

Another aspect of the invention involves a process for producing seed ofan inbred corn line including self-pollinating either of the inbredsFAR601 and FAR045, then harvesting seed. The invention also involves theseed produced by the process of inbreeding either of the inbred cornlines.

In addition, the invention involves a process for producing a hybridcorn plant utilizing at least one of the inbred corn lines FAR601 orFAR045. The process involves crossing a first inbred line with anon-identical second inbred line to produce a hybrid corn seed. Thehybrid corn seed is harvested and grown to produce a hybrid plantincluding characteristics of FAR601 or FAR045 or a combination thereof.The invention also contemplates a hybrid corn plant produced by thisprocess and the hybrid corn seed produced by the process. An example ofthis was produced by crossing FAR601 (e.g., as a male) with FAR045(e.g., as a female) to yield a hybrid RC701.

An important aspect of the inbred and hybrid corn plants associated withthe present invention has to do with the inherent stable colorexpression present in the plant parts and tissues such as the husk,inflorescences, stem and leaves which differ dramatically from those ofstandard yellow or white corn. These tissues, especially afterflowering, have a deep red to purple color which is also expressedinside the stem tissue. Even the crown roots show a much higher contentof anthocyanins as compared to standard corn varieties. The husk leavessurrounding the ear also appear different in phenotype and anthocyaninconcentration not found in the husks of standard or regular corn. Thisis quite visible. This phenomenon is characterized by a red to purplecolor which intensifies after anthesis. Accordingly, intensepigmentation is present in tissues of the inbreds and hybrid of thisinvention such as, without limitation, inflorescences, husks, cobs,stems, and grain. Consequently, the stover and grain of the inbreds ofthis invention and their hybrid have economic value above that normallypresent due to the intense pigmentation present. The deep colorationmanifested by the inbreds of this invention and their hybrid isindicative of high concentrations of pigments, such as anthocyanins, inthese tissues.

In the same manner, the grain expresses a deep red to purple colornoticeable about two to three weeks after flowering. At physiologicalmaturity, all grains are a complete deep red to purple hue. The cob ofthese inbreds and their hybrid shows a very deep red to pink and purplephenotypic expression which reaches its highest concentration when thegrain is ready for harvest.

The FAR red corn inbreds and hybrids in other respects perform in amanner similar to other standard corn hybrids grown in a field. Theseother aspects have been found to be acceptable.

All publications cited herein are hereby incorporated by reference.

DETAILED DESCRIPTION

Consumers have become increasingly concerned by the use of artificialfood colorants and, consequently, are becoming receptive to the use offood colorants extracted from plants. One plant species offering promisein terms of quantity (or yield) and quality (various colors) of suchcolorants is maize. Accordingly, the inbreds of this invention combinedesirable and productive agronomic traits with high yields ofextractible pigments.

1. INBRED LINES AND HYBRID VARIETIES

The ultimate purpose for developing maize inbred lines is to be able todependably produce hybrids. Commercially viable maize hybrids, likehybrids in many other crop species, manifest heterosis or hybrid vigorfor most economically important traits.

Plants resulting from self-pollination (or from other forms ofinbreeding) for several generations are termed inbreds (inbred lines).These inbreds are homozygous at almost all loci. When self-pollinated,these inbreds produce a genetically uniform population of true breedinginbred progeny. These inbred progeny possess genotypes and phenotypesessentially identical to that of their inbred parent. A cross betweentwo different inbreds produces a genetically uniform population ofhybrid F₁ plants. These F₁ plants are genetically uniform, but arehighly heterozygous. Progeny from a cross between two hybrid F₁ plantsare also highly heterozygous, but are not genetically uniform.

One important result of this phenomenon is that seed supplies of aninbred may be increased by self-pollinating the inbred plants.Equivalently, seed supplies of the inbred may be increased by growinginbred plants such that only pollen from these inbred plants is presentduring flowering (anthesis), e.g., in spaced or timed isolation. Seedarising from inbred parents successfully grown in isolation isgenetically identical to the inbred parents. Another important result isthat hybrids of inbred lines always have the same appearance anduniformity and can be produced by crossing the same set of inbredswhenever desired. This is because inbreds, themselves, are geneticallyuniform. Thus, a hybrid created by crossing a defined set of inbredswill always be the same. Moreover, once the inbreds giving rise to asuperior hybrid are identified, a continual supply of the hybrid seedcan be produced by crossing these identified inbred parents.

Types of hybrids include single-cross, three-way, and double-cross.Single-cross hybrids are the F₁ progeny of a cross between two inbredlines (inbreds), e.g., A*B, in which A and B are inbreds. Three-wayhybrids are the first generation progeny of a cross between asingle-cross hybrid typically used as the female and an inbred, e.g.,A*B)(C, in which A*B is a single-cross hybrid of inbreds A and B and Cis another inbred. Double-cross hybrids are the first generation progenyof a cross between two single-cross hybrids, e.g., A*B)(C*D, in whichA*B and C*D are single-cross hybrids of inbreds A and B and C and D,respectively. In the U.S., single-cross hybrids currently occupy thelargest proportion of the acreage used in maize production. As will beshown below, maize inbreds are assemblages of true breeding, homozygous,substantially identical (homogeneous) individuals. Single-cross hybridsare both homogeneous and highly heterozygous and are not true breeding.Three-way and double-cross hybrids are less homogeneous, but arenonetheless highly heterozygous and not true breeding as well. Hence,the only way of improving hybrids is improving component inbredsthereof. Improving maize inbreds involves procedures and conceptsdeveloped from the discipline of plant breeding.

2. PLANT BREEDING

Developing improved maize hybrids requires the development of improvedmaize inbreds. Maize breeding programs typically combine the geneticbackgrounds from two or more inbred lines or various other broad basedgermplasm sources into breeding populations from which new inbred linesare developed by self-pollination (or other forms of inbreeding) andselection for desired phenotypes. The newly developed inbreds arecrossed to other inbred tester lines and the hybrids from these testercrosses are then evaluated to determine whether these hybrids might havecommercial potential. Thus, the invention of a new maize varietyrequires a number of steps. As a nonlimiting illustration, these stepsmay include:

A. selecting plants for initial crosses;

B. crossing the selected plants in a mating scheme to generate F₁progeny;

C. self-pollinating the F₁ progeny to generate segregating F₂ progeny;

D. sequentially self-pollinating and selecting progeny from the F₂plants for several generations to produce a series of newly developedinbreds, which breed true and are highly uniform, yet which differ fromeach other;

E. crossing the newly developed inbred lines with other unrelated inbredlines (testers) to produce hybrid seed; and

F. evaluating the tester hybrids in replicated and unreplicatedperformance trials to determine their commercial value.

Plants are selected from germplasm pools to improve hybrid traits suchas grain and stover yield, resistance or tolerance to diseases, insects,heat and drought, stalk quality, ear retention, and end use qualities,such as quantity and quality of pigmentation. The selected plants fromthe germplasm pools are then crossed to produce F₁ plants and the F₁plants are self-pollinated to generate populations of F₂ plants.Self-pollination and selection in F₂ plants and subsequent generationsare illustrated below in a nonlimiting example of a pedigree method ofbreeding.

In the nursery, F₂ plants are self-pollinated and selected for stalkquality, reaction to diseases and insects, and other traits such aspigmentation quantity and quality, which are visually scored. During thenext growing season, seeds from each selected self-pollinated F₂ plantare planted in a row and grown as F₂-derived, F₃ families. Selection andself-pollination is practiced among and within these F₃ families. In asubsequent growing season, seeds from each selected F₃ plant are plantedin a row and grown as F₃-derived, F₄ families. Selection andself-pollination are again practiced among and within these F₄ families.In a subsequent growing season, seeds from each selected F₄ plant areplanted in a row and grown as F₄-derived, F₅ families. At this point,selection is practiced predominantly among families, rather than withinfamilies, because plants within families tend to be uniform and areapproaching homozygosity and homogeneity. Seeds from selected F₅ plantsare harvested to be further selected for uniformity prior to beingincreased.

Simultaneous with self-pollination and selection, seeds from eachselected F₂, F₃, F₄, and F₅ plant are planted in a female row in one ormore isolation blocks along with rows planted with seed of a tester(male) inbred. These isolation blocks are often grown at winterlocations so the seed harvested therefrom can be grown in performancetrials during the next growing season. Prior to anthesis, tassels fromthe selected F₂, F₃, F₄, and F₅ female plants are removed before theyshed pollen, so that the only pollen present in the isolation block isfrom the tester inbred. Seeds arising from the selected F₂, F₃, F₄, andF₅ female plants are hybrid seeds having the selected F₂, F₃, F₄, and F₅plants as maternal (seed) parents and the tester inbred as the paternal(pollen) parent.

Hybrid seeds from the isolation blocks, check hybrids, and commerciallysignificant hybrids of the same maturity are grown in replicatedperformance trials at a series of locations. Each check hybrid is theresult of crossing the tester parent and an inbred parent of knownmaturity and proven agronomic value. During the growing season, thehybrids are visually scored for any of the above-described traits. Atmaturity, plots in these trials are usually scored for the percentage ofplants with broken or tilted stalks and dropped ears. At harvest, grainyield, grain moisture, pigmentation traits, and grain test weight may bedetermined. The resulting data from these performance trials areanalyzed by calculating means and other statistics. These otherstatistics (e.g., coefficients of variation, repeatability) provideindications of the reliability (precision) of the means obtained fromthe performance trials. Means from these performance trials are thenused to further cull plants in the nursery on the basis ofunsatisfactory performance of their hybrids. Performance trials forearlier generations typically evaluate more hybrids and are planted atfewer locations than performance trials for later generations. At somepoint, seed supplies of elite inbred candidates from the nursery areincreased and are used to produce larger amounts of experimentalhybrids. These experimental hybrids are evaluated in replicatedperformance trials at maximum possible numbers of locations and may begrown alongside commercial hybrids from other seed companies in farmerfields in unreplicated trials as well. If the experimental hybridsperform well with respect to the commercial hybrids in these replicatedand unreplicated trials, they are commercialized.

While the above-described pedigree method is widely used to developmaize inbreds, variations are widely used as well. Moreover, otherbreeding method protocols such as those for bulks, backcrossing,recurrent selection, and mass selection may be practiced in addition to,or in lieu of, the pedigree method described above. Theories andexemplary protocols for the pedigree method, bulk method, recurrentselection, and mass selection are known to the art, but are disclosedin, e.g., A. R. Hallauer and J. B. Miranda Fo, Quantitative Genetics inMaize Breeding, Iowa State University Press, Ames, Iowa (1981); G.Namkoong, Introduction to Quantitative Genetics in Forestry, U.S. Dept.Agric. Forest Service Tech. Bull. No. 1588 (1979); F. N. Briggs and P.F. Knowles, Introduction to Plant Breeding, Reinhold Publishing Company,New York (1967), R. W. Allard, Principles of Plant Breeding, Wiley andSons, New York (1960), N. W. Simmonds, Principles of Crop Improvement,Longman Group, Ltd., London (1979); and J. M. Poehlman, Breeding FieldCrops, 2d Ed., AVI Publishing Co., Inc. Westport, Conn. (1979).

As discussed above, hybrids of promising advanced breeding lines arethoroughly tested and compared to appropriate check hybrids inenvironments representative of the commercial target area(s), usuallyfor 2-3 years. The best hybrids identified by these performance trialsare candidates for commercial exploitation. Seed of each of the newlydeveloped inbred parents of these hybrids is further purified andincreased in steps leading to commercial production. These prerequisiteactivities to marketing newly developed hybrids usually take from eightto 12 years from the time the first breeding cross is made. Therefore,development of new cultivars is a time-consuming process requiringprecise planning and efficient allocation and utilization of limitingresources.

Identification of genetically superior individuals is one of the mostchallenging issues confronting the plant breeder. For many economicallyimportant traits, the true genotypic expression of the trait is maskedby effects of other (confounding) plant traits and environmentalfactors. One method of identifying a superior hybrid is to observe itsperformance relative to other experimental hybrids and to a series ofwidely grown standard cultivars. However, because a single observationis usually inconclusive, replicated observations over a series ofenvironments are necessary to provide an estimate of the genetic worthof a hybrid.

Maize is an important and valuable field crop. Hence, a continuing goalof plant breeders is to develop high-yielding maize hybrids possessingcommercially acceptable quantities of desirable pigments, which areotherwise agronomically desirable and which are produced by stableinbred lines. To accomplish this goal, the maize breeder mustcontinually develop superior inbred parent lines. Developing superiorinbred parent lines requires identification and selection of geneticallyunique, superior individuals from within segregating populations.

Each segregating population is the result of a combination of amultitude of genetic crossover events, independent assortment ofspecific combinations of alleles at many gene loci, and inheritance oflarge groups of genes together due to the effects of linkage. Thus, theprobability of selecting any single individual with a specific superiorgenotype from a breeding cross is infinitesimally small due to the largenumber of segregating genes and the virtually unlimited recombinationsof these genes. Nonetheless, the genetic variation present among thesegregating progeny of a breeding cross enables the identification ofrare and valuable new genotypes. These rare and valuable new genotypesare neither predictable nor incremental in value, but are rather theresult of expressed genetic variation. Thus, even if the genotypes ofthe parents of the breeding cross can be completely characterized and adesired genotype known, only a few, if any, individuals with the desiredgenotype may be found within a large, segregating F₂ population.Typically, however, neither the genotypes of the parents used in thebreeding cross nor the desired progeny genotype to be selected are knownto any extent.

In addition to the preceding problem, it is not known with any degree ofcertainty how the new genotype would interact with the environment. Thisuncertainty is measured statistically by genotype-by-environmentinteractions and is an important, yet unpredictable, factor in plantbreeding. A breeder of ordinary skill in the art can neither predict norcharacterize a priori a new desirable genotype, how the genotype willinteract with various climatic factors, or the resulting phenotypes ofthe developing lines, except perhaps in a very broad and gross fashion.A breeder of ordinary skill in the art would also be unable to re-createthe same line twice from the same original parents, because the breederis unable to direct how the parental genomes will recombine in theprogeny or how the resulting progeny will interact with environmentalconditions when undergoing selection. This unpredictability results inthe expenditure of large amounts of limited research resources todevelop each superior new maize inbred line.

A reliable method of controlling male fertility (pollen viability) inplants provides means for efficient and economical subsequent hybridproduction. This is also the case when plant breeders are developingmaize hybrids in breeding programs. All breeding programs rely on somesort of system or method of pollen control and there are several methodsof pollen control available to breeders. These pollen control methodsinclude barriers such as bags for covering silks and collecting pollenfrom individual plants, manual or mechanical emasculation (detasseling),cytoplasmic male-sterility (CMS), genetic male-sterility, andgametocides.

Hybrid maize seed is usually produced commercially by using amale-sterility system, manual or mechanical detasseling, or acombination of both. In typical commercial hybrid seed production,alternate strips of two maize inbreds are planted in a field. Thetassels are removed from the inbred designated to be the seed or femaleparent. Alternatively, the female is male-sterile and is not detasseled.If there is sufficient isolation from sources of foreign maize pollen,the ears of the female inbred will be fertilized only with pollen fromthe other (male) inbred. The resulting seed, harvested from the femaleparents in a successful hybrid production effort, is hybrid F₁ seed,which will germinate and grow into hybrid F₁ plants.

Manual or mechanical detasseling can be avoided by using inbreds withcytoplasmic male-sterility (CMS). CMS requires both a homozygous nuclearlocus and the presence of a cytoplasmic factor for sterility. Otherwise,the plant will produce viable pollen. The CMS system requires A-lines(females), B-lines (maintainers), and R-lines (males). Male-sterileA-lines are homozygous for a nuclear allele for pollen sterility andpossess the cytoplasmic factor for pollen sterility as well. B-linesproduce viable pollen because they are homozygous for the sterilenuclear allele, but possess a fertile cytoplasmic factor. With theexception for the allele for pollen fertility, B-lines usually have anuclear genome essentially identical to their complimentary A-line.R-lines are homozygous for a nuclear allele for fertility and possess afertile cytoplasmic factor. Thus, R-lines produce viable pollen. Seed ofmale-sterile A-lines is increased by being pollinated by complimentaryB-lines. The resulting seed grows into male-sterile A-line plantsbecause the fertile cytoplasmic factor from the B-lines is nottransmitted by B-line pollen. Hybrid seed is produced by pollinatingA-line plants with pollen from R-line plants. The resulting hybrid seedis heterozygous at the nuclear locus and possesses the sterilecytoplasmic factor. Thus, the hybrid seed will grow into plants whichproduce viable pollen.

In addition to CMS, there are several methods conferring geneticmale-sterility. One method involves multiple loci (including a markergene in one case) which confer male-sterility, as disclosed in U.S. Pat.Nos. 4,654,465 and 4,727,219 to Brar et al. Another method disclosed byU.S. Pat. Nos. 3,861,709 and 3,710,511 to Patterson uses chromosomalreciprocal translocations, deficiencies, and duplications. In additionto these methods, U.S. Pat. No. 5,432,068 to Albertsen et al., describesa system of induced nuclear male-sterility which includes: identifying agene critical to male fertility; “silencing” this critical gene;replacing the native promoter from the critical gene with an induciblepromoter; and inserting the genetically engineered gene back into theplant. The resulting plant is male-sterile while the inducible promoteris not operative because the male fertility gene is not transcribed.Fertility is restored by inducing the promoter with a non-phytotoxicchemical which induces expression of the critical gene, thereby causingthe gene conferring male fertility to be transcribed. U.S. Pat. Nos.5,689,049 and 5,689,051 to Cigan et al. disclose a transgenic maizeplant rendered male-sterile by being transformed with a geneticconstruct including regulatory elements and DNA sequences capable ofacting in a fashion to inhibit pollen formation or function.

Yet another male-sterility system delivers a gene encoding a cytotoxicsubstance into the plant. The cytotoxic substance is associated with amale tissue-specific promoter or an antisense system. In each instance,a gene critical to fertility is identified and an antisensetranscription to that gene is inserted in the plant (see e.g.,Fabinjanski, et al., EPO 89/3010153.8 Publication No. 329,308 and PCTApplication No. PCT/CA90/00037, published as WO 90/08828.

Another system potentially useful to confer male-sterility usesgametocides. Gametocides are topically applied chemicals affecting thegrowth and development of cells critical to male fertility. Applicationof gametocides affects fertility in the plants only for the growingseason in which the gametocide is applied. See, e.g., U.S. Pat. No.4,936,904 to Carlson (N-alkyl-2-aryl-4-oxonicotinates,N-alkyl-5-aryl-4-oxonicotinates, N-alkyl-6-aryl-4-oxonicotinates,N-alkyl-2,6-diaryl-4-oxonicotinates). Inbred genotypes differ in theextent to which they are rendered male-sterile by gametocides and in thegrowth stages at which the gametocides must be applied.

During hybrid seed production, incomplete detasseling or incompleteinactivation of pollen from the female parent will cause some of thefemale parent plants to be self-pollinated. These selfed female plantswill produce seed of the female inbred, rather than the desired hybridseed. The selfed seed of the female plants will then be harvested andpackaged along with the hybrid seed. Alternatively, seed from the maleinbred line may also be present among hybrid seed if the male plants arenot eliminated after pollination. In either case, once the mixture ofhybrid and “selfed” seed is planted it is possible to identify andselect the female or male inbreds growing among hybrid plants. Typicallythese “selfs” are easily identified and selected because of theirdecreased vigor for vegetative and/or reproductive characteristics(e.g., shorter plant height, small ear size, ear and kernel shape, orcob color). Identification of these selfs can also be accomplishedthrough molecular marker analyses. See, e.g., Smith et al., “TheIdentification of Female Selfs in Hybrid Maize: A Comparison UsingElectrophoresis and Morphology”, Seed Science and Technology 14:1-8(1995). Through these technologies, the homozygosity of theself-pollinated line can be verified by analyzing allelic composition atvarious loci along the genome. These methods allow for rapididentification of the invention disclosed herein. See also, Sarca etal., “Identification of Atypical Plants in Hybrid Maize Seed byPostcontrol and Electrophoresis,” Probleme de Genetica Teoritica siAplicata Vol. 20(1): 29-42. As is apparent to one skilled in the art,the foregoing are only some of the ways by which an inbred can beobtained and seed supplies of inbreds and hybrids increased.

3. GRAIN AND STOVER PRODUCTION

This invention is contemplated to include producing stover and grainwhen hybrids with FAR601 or FAR045 as a parent are grown. Typically seedof these hybrids is planted in soil with adequate moisture to supportgermination, emergence, and subsequent growth and development.Alternatively, soil moisture is added by irrigation. Normal culturalpractices to achieve proper soil fertility and manage weeds, insects,and diseases may be undertaken during the growing season as necessary.These cultural practices are known to persons of skill in the art andvary widely according to particular geographic regions, growerpreferences, and economic considerations. The corn plants may be choppedfor silage, typically when the developing grain is at the half-milkstage. The grain is harvested when physiologically mature, usually withcombines, then dried to a moisture content sufficiently low for storage.The grain may then be used for feed, food, and industrial purposes,examples of which are disclosed herein.

4. DERIVATION

This invention is considered to include processes of developing derived(introgressed) maize inbred lines and plants, seeds, and parts resultingthereof. Processes of developing derived inbred lines include thoseprocesses, wherein single genes or alleles or some small plurality ofgenes or alleles are introgressed into FAR601 or FAR045, resulting in aderived inbred which expresses the introgressed gene(s) or allele(s)(i.e. trait(s)), but otherwise retains the phenotype and genotype ofFAR601 or FAR045 described herein. Examples of introgressed genes oralleles include insect or disease resistance, genes from other maizeplants, or alleles or genes originating from other species. Non-limitingexamples of these genes or alleles are disclosed in Coe et al., “TheGenetics of Corn,” IN Corn and Corn Improvement, G. F. Sprague and J. W.Dudley, Editors, American Society of Agronomy, Madison, Wis. (1988).Other nonlimiting examples of genes or alleles which might beintrogressed into the present invention are disclosed hereinbelow.Methods of introgression may include such protocols as backcrossing,tissue culture to induce somoclonal variation, impaling plant cells withneedle-like bodies, use of indeterminate gametophyte, anther culture,and transformation.

Backcrossing protocols are disclosed, e.g. in above-referenced F. N.Briggs and P. F. Knowles, Introduction to Plant Breeding, ReinholdPublishing Company, New York (1967), R. W. Allard, Principles of PlantBreeding, Wiley and Sons, New York (1960), N. W. Simmonds, Principles ofCrop Improvement, Longman Group, Ltd., London (1979); and J. M.Poehlman, Breeding Field Crops, 2d Ed., AVI Publishing Co., Inc.Westport, Conn. (1979). Use of indeterminate gametophyte-facilitated(ig1) introgression of cytologically inherited traits is disclosed by,e.g., J. L. Kermicle, “Androgenesis Conditioned by a Mutation in Maize,”Science 166: 1422-1424 (1969).

Isolated microspore, anther culture and regeneration of fertile maizeplants are disclosed in U.S. Pat. No. 5,445,961 to Genovesi et al.Introgression protocols using anther culture are disclosed, e.g., byBarnabas et al., “Ultrastructural Studies on Pollen Embryogenesis inMaize (Zea mays L)”, Plant Cell Rep. 6: 212-215 (1987); Dieu et al.,“Further Studies of Androgenetic Embryo Production and PlantRegeneration From In Vitro Cultured Anthers in Maze (Zea mays L.),”Maydica 31: 245-259 (1986); Pace et al., “Anther Culture of Maize andthe Visualization of Embryogenic Microspores by Fluorescent Microscopy,”Theor. Appl. Genet. 73: 863-869 (1987); Petolino et al., “Anther Cultureof Elite Genotypes of Maize,” Crop Sci. 26: 1072-1074 (1986); and Tsayet al., “Factors Affecting Haploid Plant Regeneration from Maize AntherCulture,” J. Plant Physiol. 126: 33-40 (1986).

Exemplary transformation protocols are disclosed, e.g., by U.S. Pat. No.5,302,523 to Coffee et al. (transformed maize via needle-like bodies);U.S. Pat. No. 5,384,253 to Krzyzek et al. (electroporation); U.S. Pat.No. 5,371,003 to Murray et al. (transformation via tissues withinhorizontal electrophoresis gel in the presence of non-pulsed electriccurrent); U.S. Pat. No. 5,591,616 to Hiei et al. (Agrobacterium-mediatedtransformation); U.S. Pat. No. 5,569,597 to Grimsley et al.(Agrobacterium-mediated maize transformation); U.S. Pat. No. 5,877,023to Sautter et al. (microprojectile-facilitated transformation); U.S.Pat. No. 5,736,369 to Bowen et al. (microprojectile-facilitatedtransformation); U.S. Pat. Nos. 5,886,244 and 5,990,387 to Tomes et al.(microprojectile-facilitated transformation); U.S. Pat. No. 5,776,900 toShillito et al. (regeneration of maize protoplasts transformed withelectroporation and polyethylene glycol (PEG)); U.S. Pat. Nos. 5,767,367and 5,792,936 to Dudits et al. (regeneration of PEG-transformedprotoplasts of auxin-autotrophic maize genotype); U.S. Pat. Nos.5,780,708 and 5,990,390 to Lundquist et al. (fertile,microprojectile-facilitated transgenic maize plants expressing dalaponresistance); U.S. Pat. Nos. 5,780,709 and 5,919,675 to Adams et al.(microprojectile- and electroporation-facilitated maize transformants);U.S. Pat. No. 5,932,782 to Bidney (microprojectile-deliveredAgrobacterium); U.S. Pat. No. 5,981,840 to Zhao et al.(Agrobacterium-transformed maize); and U.S. Pat. No. 5,994,624 toTrolinder et al. (maize transformation via recombinant Agrobacterium DNAinjected into plant tissues via needleless injection device). Anexemplary transformation protocol is more fully disclosed hereinbelow.

5. FURTHER USES

This invention is also contemplated to include processes or methods ofproducing a maize plant by crossing a first parent maize plant with asecond parent maize plant in which the first or second parent maizeplant is the inbred maize line FAR601 or FAR045. Moreover, both thefirst and second parent maize plants may include the inbred maize linesFAR601 and/or FAR045.

This invention is also directed to processes or methods of producingFAR601 or FAR045-derived maize plant or an inbred maize plant withFAR601 or FAR045 as a parent in at least one of the initial breedingcrosses accomplished by crossing inbred maize line FAR601 or FAR045 witha second maize plant and growing the progeny seed. The method mayfurther include repeating crossing and growing the FAR601-derived orFAR045-derived plant until the substantial genotype of FAR601 or FAR045is recovered. Thus, any methods using the inbred maize line FAR601 orFAR045 are contemplated to be within the scope of this invention, e.g.,selfing, backcrossing, hybrid production, crosses to other hybrids,inbreds, populations, and the like. All plants produced using inbredmaize line FAR601 or FAR045 as a parent are contemplated to be withinthe scope of this invention, including plants derived from inbred maizeline FAR601 or FAR045. It should be further understood that inbred maizeline FAR601 or FAR045 can, through routine manipulation known to skilledpersons in the art, be produced in a male-sterile form and that suchembodiments are contemplated to be within the scope of the presentinvention as well.

As used herein, the term “plant” includes whole or entire plants andparts thereof. Such exemplary plant parts may include plant cells, plantprotoplasts, plant cell tissue cultures, plant calli, plant clumps,plant cell suspension cultures, and plant protoplasts. Also includedwithin the definition of the term “plant” are plant cells present inplants or parts of plants, e.g., zygotes, embryos, embryonic organs,pollen, ovules, flowers, seeds, ears, cobs, leaves, husks, stalks,roots, root tips, anthers, and silks.

Tissue Culture of Maize

Regeneration of maize plants by tissue culture methods is now anexercise requiring only routine experimentation to a person skilled inthe art. For example, Duncan et al. (Planta 165:322-332 (1985)) reported97% of the plant genotypes cultured produced calli capable of plantregeneration. Plants were regenerated from 91% of the calli from anotherset of inbreds and hybrids in a subsequent experiment.

Songstad et al., (Plant Cell Reports, 7:262-265 (1988)) reported severalmedia additions enhancing regenerability of callus of two inbred lines.Other published reports also indicated “nontraditional” tissues capableof producing somatic embryogenesis and plant regeneration. For example,Rao, et al. (Maize Genetics Cooperation Newsletter, 60:64-65 (1986))reported somatic embryogenesis from glume callus cultures. Conger, etal. (Plant Cell Reports, 6:345-347 (1987)) reported somaticembryogenesis from tissue cultures of maize leaf segments. Thus, it isclear that the state of the art is such that these tissue culturemethods of obtaining regenerated plants are routinely used with veryhigh rates of success.

Maize tissue culture is described generally in European PatentApplication, Publication 160,390 and with respect to inbred line B73 inU.S. Pat. No. 5,134,074 to Gordon et al. Maize tissue culture proceduresare also described by U.S. Pat. No. 4,581,847 to Hibberd et al., by Kamoet al. “Establishment and Characterization of Long-Term Embryonic MaizeCallus and Cell Suspension Cultures,” Plant Science 45: 111-117, byVasil et al., “Plant Regeneration from Friable Embryonic Callus and CellSuspension Culture of Zea mays L.,” J. Plant Physiol. 124:399-408(1986), by Green et al., “Plant Regeneration in Tissue Culture ofMaize,” Maize for Biological Research (Plant Molecular BiologyAssociation, Charlottesville, Va. 1982, at 367-372) and by Duncan, etal., “The Production of Callus Capable of Plant Regeneration fromImmature Embryos of Numerous Zea Mays Genotypes,” 165 Planta 322-332(1985). Thus, another aspect of this invention is to provide cells,which undergo growth and differentiation and subsequently produce maizeplants with the physiological and morphological characteristics ofinbred maize line FAR601 or FAR045.

Somaclonal variation within inbred lines which have undergone tissueculture and regeneration have been reported by Edallo et al.(“Chromosome Variation and Frequency of Spontaneous Mutants AssociatedWith In Vitro Culture and Plant Regeneration in Maize,” Maydica 26:39-56 (1981)); McCoy et al. (“Chromosome Stability in Maize (Zea MaysL.) Tissue Culture and Sectoring in Some Regenerated Plants,” Can. J.Genet. Cytol. 24: 559-565 (1982)), Earle et al. (“Somaclonal Variationin Progeny of Plants From Corn Tissue Culture,” pp 139-152, IN R. R.Henke et al. (ED.) Tissue Culture in Forestry and Agriculture, PlenumPress, N.Y. (1985)); and Lee et al. (“Agronomic Evaluation of InbredLines Derived From Tissue Cultures of Maize,” Theor. Appl. Genet. 75:841-849 (1988)). Hence, genetic variation and derived lines may bedeveloped from this invention by tissue culture protocols.

The utility of inbred maize line FAR601 or FAR045 also extends tocrosses with other species. Suitable species will be of the familyGramineae, and especially genera such as Zea, Tripsacum, Coix,Schlerachne, Polytoca, Chionachne, and Trilobachne, of the tribeMaydeae. Potentially suitable for crosses with inbred maize line FAR601or FAR045 may be the various varieties of grain sorghum, Sorghum bicolor(L.) Moench. or other species within the genus Sorghum.

6. TRANSFORMATION

Molecular biological techniques now allow genes encoding specificprotein products to be isolated and characterized. It has long beenviewed as advantageous to modify maize plant genomes to contain andexpress foreign genes, or additional, or modified versions of native orendogenous genes (perhaps driven by different promoters) to alter traitsof a plant in a specific, directed manner. Such foreign, additionaland/or modified genes are referred to herein collectively as“transgenes” and several methods for producing transgenic plants havebeen developed. Accordingly, embodiments of this invention also includederived inbreds which are transformed versions of inbred maize lineFAR601 or FAR045.

Plant transformation requires construction of an expression vector tofunction in plant cells. Such an expression vector includes DNA. Thevector DNA, in turn, includes a gene under control of, or operativelylinked to, a regulatory element such as a promoter. The expressionvector may contain one or more such operably linked gene/regulatoryelement combinations; may be in the form of a plasmid; and can also beused alone, or in combination with other plasmids, to transform maizeplants using transformation methods such as those described below.

7. MARKER GENES

Expression vectors usually include at least one genetic marker operablylinked to a regulatory element such as a promoter. The regulatoryelement allows transformed cells containing the marker to be recoveredeither by negative or positive selection. Negative selection includesinhibiting the growth of cells not containing the selectable markergene. By contrast, positive selection includes screening for the productencoded by the genetic marker. Many commonly used selectable markers foridentifying transformed plant cells are known in the art. Such exemplaryselectable markers include genes encoding enzymes which metabolicallydetoxify a selective chemical agent such as an antibiotic or anherbicide. Other selectable markers include genes encoding an alteredtarget which is insensitive to an inhibitor. A few positive selectionmethods are also known.

One commonly used selectable marker is the neomycin phosphotransferaseII gene (nptII), isolated from transposon Tn5 and conferring resistanceto kanamycin. Fraley et al., Proc. Natl. Acad. Sci. U.S.A., 80: 4803(1983); U.S. Pat. No. 5,858,742 to Fraley et al. Another commonly usedselectable marker gene is the hygromycin phosphotransferase geneconferring resistance to the antibiotic hygromycin. Vanden Elzen et al.,Plant Mol. Biol., 5: 299 (1985).

Other selectable marker genes of bacterial origin conferring resistanceto antibiotics include gentamycin acetyl transferase, streptomycinphosphotransferase, aminoglycoside-3′-adenyl transferase, and bleomycinresistance determinant. Hayford et al., Plant Physiol. 86: 1216 (1988);Jones et al., Mol. Gen. Genet., 210: 86 (1987); Svab et al., Plant Mol.Biol. 14: 197 (1990); and Hille et al., Plant Mol. Biol. 7: 171 (1986).

Still other selectable markers confer resistance to herbicides such asglyphosate, glufosinate, or bromoxynil. Comai et al., Nature 317:741-744 (1985); Gordon-Kamm et al., Plant Cell 2: 603-618 (1990); andStalker et al., Science 242: 419-423 (1988).

Yet other selectable marker genes include mouse dihydrofolate reductase,plant 5-enolpyruvylshikimate-3-phosphate synthase, and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13: 67(1987); Shah et al., Science 233: 478 (1986); and Charest et al., PlantCell Rep. 8: 643 (1990).

Another class of marker genes useful in plant transformation requiresscreening putatively transformed plant cells, rather than direct geneticselection of transformed cells. These genes are used to quantify orvisualize spatial patterns of gene expression in specific tissues.Marker genes of this nature are frequently termed “reporter genes”because they can be fused to a gene or gene regulatory sequence toinvestigate gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase and chloramphenicol acetyltransferase.Jefferson, Plant Mol. Biol. Rep. 5: 387 (1987); Teeri et al., EMBO J. 8:343 (1989); Koncz et al., Proc. Natl. Acad. Sci. U.S.A. 84: 131 (1987);and De Block et al., EMBO J. 3: 1681 (1984). Until recently, methods forvisualizing GUS activity required destruction of the living plantmaterial. However, in vivo methods for visualizing GUS activity notrequiring destruction of plant tissue are now available. MolecularProbes Publication 2908, Imagene Green™, p. 1-4 (1993); and Naleway etal., J. Cell Biol. 115: 151a(1991).

Another method of identifying rare transformation events includes usinga gene encoding a dominant constitutive regulator of the Zea maysanthocyanin pigmentation pathway. Ludwig et al., Science 247: 449(1990). A gene encoding for Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263: 802 (1994).

8. PROMOTERS

Genes in expression vectors must be driven by a nucleotide sequencecomprising a regulatory element such as a promoter. Several types ofpromoters are now known, as are other regulatory elements which can beused singly or in combination with promoters. As used herein “promoter”includes a region of DNA upstream from the initial site oftranscription. The promoter is involved in recognizing and binding RNApolymerase and other proteins during transcription initiation. A “plantpromoter” is a promoter capable of initiating transcription in plantcells.

Examples of promoters under developmental control include promoterswhich preferentially initiate transcription in certain tissues, such asleaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma.Such promoters are referred to as “tissue-preferred.” Promotersinitiating transcription only in certain tissues are referred to as“tissue-specific.” A “cell type-specific” promoter primarily drivesexpression only in certain cell types present in specific organs, e.g.,vascular cells in roots or leaves. An “inducible” promoter is a promoterunder environmental control. Examples of environmental conditionsaffecting transcription by inducible promoters include anaerobicconditions or the presence of light. Tissue-specific, tissue-preferred,cell type-specific, and inducible promoters constitute the class of“non-constitutive” promoters. In contrast to non-constitutive promoters,“constitutive” promoters function under most environmental conditions.

A. Inducible Promoters

An inducible promoter may be operably linked to a gene to be expressedin maize. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence. The signal sequence, inturn, is operably linked to a gene to be expressed in maize. With aninducible promoter, the rate of transcription increases in response toan inducing agent.

Any inducible promoter can be used in conjunction with this invention.See, e.g., Ward et al. Plant Mol. Biol. 22: 361-366 (1993). Exemplaryinducible promoters include, but are not limited to, the promoter theACEI system responding to copper (Mett et al. PNAS 90: 4567-4571(1993)); the maize In2 gene responding to benzenesulfonamide herbicidesafeners (Hershey et al., Mol. Gen. Genetics 227: 229-237 (1991) andGatz et al., Mol. Gen. Genetics 243: 32-38 (1994)); or the Tet repressorfrom Tn10 (Gatz et al., Mol. Gen. Genet. 227: 229-237 (1991). Onesuitable inducible promoter responds to an inducing agent to whichplants do not normally respond. One such exemplary inducible promoter isinduced by a glucocorticosteroid hormone. Schena et al., Proc. Natl.Acad. Sci. U.S.A. 88: 0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene to be expressed inmaize. Alternatively, the constitutive promoter is operably linked to anucleotide sequence encoding a signal sequence which, in turn, isoperably linked to a gene to be expressed in maize. Many differentconstitutive promoters can be utilized with respect to the inbred ofthis invention. Exemplary constitutive promoters include, but are notlimited to, promoters from plant viruses such as the 35S promoter fromCaMV (Odell et al., Nature 313: 810-812 (1985); U.S. Pat. No. 5,858,742to Fraley et al.); promoters from such plant genes as rice actin(McElroy et al., Plant Cell 2: 163-171 (1990)); ubiquitin (Christensenet al., Plant Mol. Biol. 12: 619-632 (1989) and Christensen et al.,Plant Mol. Biol. 18: 675-689 (1992)); pEMU (Last et al., Theor. Appl.Genet. 81: 581-588 (1991)); MAS (Velten et al., EMBO J. 3: 2723-2730(1984)) and maize H3 histone (Lepetit et al., Mol. Gen. Genet. 231:276-285 (1992) and Atanassova et al., Plant Journal 2(3): 291-300(1992)); and the ALS promoter, a XbaI/NcoI fragment 5′ to the Brassicanapus ALS3 structural gene or a nucleotide sequence with substantialsequence similarity (PCT Application No. WO96/30530).

C. Tissue-Specific or Tissue-Preferred Promoters

A tissue-specific promoter is operably linked to a gene to be expressedin maize. Optionally, the tissue-specific promoter is operably linked toa nucleotide sequence encoding a signal sequence which is operablylinked to a gene to be expressed in maize. Plants transformed with agene operably linked to a tissue-specific promoter produce the proteinproduct of the transgene exclusively, or preferentially, in a specifictissue.

Any tissue-specific or tissue-preferred promoter can be introgressedinto the inbred of this invention. Exemplary tissue-specific ortissue-preferred promoters include, but are not limited to, aroot-preferred promoter, such as that from the phaseolin gene (Murai etal., Science 23: 476-482 (1983) and Sengupta-Gopalan et al., Proc. Natl.Acad. Sci. USA 82: 3320-3324 (1985)); a leaf-specific and light-inducedpromoter such as that from cab or rubisco (Simpson et al., EMBO J.4(11): 2723-2729 (1985) and Timko et al., Nature 318: 579-582 (1985));an anther-specific promoter such as that from LAT52 (Twell et al., Mol.Gen. Genet. 217: 240-245 (1989)); a pollen-specific promoter such asthat from Zml3 (Guerrero et al., Mol. Gen. Genet. 224: 161-168 (1993));and a microspore-preferred promoter such as that from apg (Twell et al.,Sex. Plant Reprod. 6: 217-224 (1993)).

9. SIGNAL SEQUENCES FOR TARGETING PROTEINS TO SUBCELLULAR COMPARTMENTS

Proteins produced by transgenes may be transported to a subcellularlocation such as a chloroplast, vacuole, peroxisome, glyoxysome, cellwall or mitochondrion, or for secretion into the apoplast, by operablylinking the nucleotide sequence encoding a signal sequence to the 5′and/or 3′ region of a gene encoding the protein of interest. Targetingsequences at the 5′ and/or 3′ end of the structural gene may determinewhere the encoded protein is ultimately compartmentalized during proteinsynthesis and processing. The presence of a signal sequence directs apolypeptide to an intracellular organelle, a subcellular compartment, orto the apoplast for secretion. Many signal sequences are known in theart. See, e.g., Becker et al., Plant Mol. Biol. 20: 49 (1992); P. S.Close, Master's Thesis, Iowa State University (1993); Knox, et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes FromBarley,” Plant Mol. Biol. 9: 3-17 (1987); Lerner et al., Plant Physiol.91: 124-129 (1989); Fontes et al., Plant Cell 3: 483-496 (1991);Matsuoka et al., Proc. Natl. Acad. Sci. 88: 834 (1991); Gould et al., J.Cell Biol 108: 1657 (1989); Creissen et al., Plant J. 2: 129 (1991);Kalderon et al., “A short amino acid sequence able to specify nuclearlocation,” Cell 39: 499-509 (1984); and Stiefel et al., “Expression of amaize cell wall hydroxyproline-rich glycoprotein gene in early leaf androot vascular differentiation,” Plant Cell 2: 785-793 (1990).

10. FOREIGN PROTEIN GENES AND AGRONOMIC GENES

A foreign protein can be produced by transgenic plants of this inventionand may further be produced in commercial quantities. Thus, techniquesfor selection and propagation of transformed plants provide a pluralityof transgenic plants, which may be harvested in a conventional manner. Aforeign protein expressed in the transgenic plants can then be extractedeither from a specific tissue or from total harvested plant biomass.Protein extraction from plant biomass can be accomplished by methodswhich are discussed, e.g., by Heney et al., Anal. Biochem. 114: 92-96(1981).

Thus, this invention is contemplated to include transformed, thereforederived, embodiments of inbred maize line FAR601 or FAR045. In anotherembodiment, the biomass of interest is the vegetative tissue of inbredmaize line FAR601 or FAR045. In yet another embodiment, the biomass ofinterest is grain (seed). For transgenic plants, a genetic map can begenerated, primarily via conventional Restriction Fragment LengthPolymorphisms (RFLP), Polymerase Chain Reaction (PCR) analysis, RandomAmplified Polymorphic DNA (RAPD), Amplified Fragment LengthPolymorphisms (AFLP), Single Nucleotide Polymorphisms (SNP), and SimpleSequence Repeats (SSR), which identify the approximate chromosomallocation of the integrated DNA. For exemplary methodologies in thisregard, see Glick et al., Methods in Plant Molecular Biology andBiotechnology, 269-284 (CRC Press, Boca Raton, 1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa given transgenic plant. Hence, if unauthorized propagation occurs andcrosses of the present inbred are made to other germplasm, the map ofthe integration region can be compared to similar maps of suspectplants, thereby determining whether the suspect plants have a commonparentage with the subject plant. Map comparisons require hybridizationand subsequent RFLP, PCR, SSR, RAPD, AFLP, SNP and/or sequencing, all ofwhich are known techniques.

Agronomic genes can be expressed in the transformed plants of thisinvention. More particularly, plants of this invention can betransformed, or otherwise derived, to express various phenotypes ofagronomic interest. Exemplary genes implicated in this regard include,but are not limited to, those categorized below.

11. GENES CONFERRING RESISTANCE TO PESTS OR DISEASES

A. Plant Disease Resistance Genes.

Plant defenses are often activated by specific interaction between theproduct of a disease resistance gene (R) in the plant and the product ofa corresponding avirulence (Avr) gene in the pathogen. A plant varietycan be transformed with a cloned disease resistance gene to developplants resistant to pathogen strains. See, e.g., Jones et al., Science266: 789 (1994) (cloning of tomato Cf-9 gene resistant to Cladosporiumfulvum); Martin et al., Science 262: 1432 (1993) (tomato Pto generesistant to Pseudomonas syringae pv. tomato encoding a protein kinase);Mindrinos et al., Cell 78:1089 (1994) (Arabidopsis RSP2 gene resistantto Pseudomonas syringae); U.S. Pat. No. 5,789,214 to Ryals et al.(chemically regulatable DNA sequences regulating transcription ofpathogenesis-related proteins); and PCT Patent Application PublicationWO95/16776 to Putman et al. (derivatives of tachyplesin peptide withantimicrobial activity against plant pathogens).

B. Bacillus thuringiensis (B.t.) Proteins.

Bacillus thuringiensis (B.t.) proteins, derivatives thereof, or asynthetic polypeptides modeled thereon. See, e.g., Geiser et al., Gene48: 109 (1986) (cloning and nucleotide sequencing of B.t. δ-endotoxingene). DNA molecules encoding δ-endotoxin genes are designated as ATCCAccession Nos. 40098, 67136, 31995 and 31998 and can be obtained fromAmerican Type Culture Collection, Manassas, Va. 20110.

C. Lectins

Lectins. See, e.g., Van Damme et al., Plant Molec. Biol. 24: 25 (1994)(nucleotide sequences of Clivia miniata mannose-binding lectin genes).

D. Vitamin-Binding Proteins

Vitamin-binding proteins such as avidin. See, e.g., PCT Application No.US93/06487 (avidin and avidin homologues as larvicides against insectpests).

E. Enzyme Inhibitors

Enzyme inhibitors such as protease inhibitors or amylase inhibitors.See, e.g., Abe et al., J. Biol. Chem. 262: 16793 (1987) (nucleotidesequence of rice cysteine proteinase inhibitor); Huub et al., PlantMolec. Biol. 21: 985 (1993) (nucleotide sequence of cDNA encodingtobacco proteinase inhibitor I); and Sumitani et al., Biosci. Biotech.Biochem. 57: 1243 (1993) (nucleotide sequence of Streptomycesnitrosporeus α-amylase inhibitor).

F. Insect-Specific Hormone or Pheromone

An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, e.g., Hammock et al., Nature 344:458 (1990), (baculovirus expression of cloned juvenile hormone esterase,an inactivator of juvenile hormone).

G. Insect-Specific Peptides or Neuropeptides

Insect-specific peptides or neuropeptides disrupting pest physiologies.See, e.g., Regan, Biol. Chem. 269: 9 (1994) (expression cloning yieldsDNA coding for insect diuretic hormone receptor); and Pratt et al.,Biochem. Biophys. Res. Comm. 163: 1243 (1989) (allostatin identified inDiploptera puntata); U.S. Pat. No. 5,266,317 to Tomalski et al. (genesencoding insect-specific, paralytic neurotoxins).

H. Insect-Specific Venoms

Insect-specific venoms produced in nature by, e.g., snakes, wasps. See,e.g., Pang et al., Gene 116: 165 (1992) (heterologous expression inplants of a gene coding a scorpion insectotoxic peptide).

I. Enzymes

Enzymes responsible for hyperaccumulation of monterpenes,sesquiterpenes, steroids, hydroxamic acids, phenylpropanoid derivativesor other non-protein molecules with insecticidal activity.

Enzymes involved in the modification, including post-translationalmodification, of biologically active molecules. Such enzymes arecontemplated to include natural or synthetic glycolytic enzymes,proteolytic enzymes, lipolytic enzymes, nucleases, cyclases,transaminases, esterases, hydrolases, phosphatases, kinases,phosphorylases, polymerases, elastases, chitinases and glucanases. See,e.g., PCT Application No. WO 93/02197 to Scott et al. (callase genenucleotide sequence). DNA molecules containing chitinase-encodingsequences can be obtained, e.g., from the ATCC under Accession Nos.39637 and 67152. See, also Kramer et al., Insect Biochem. Molec. Biol.23: 691 (1993) (nucleotide sequence of cDNA-encoding tobacco hookwormchitinase); and Kawalleck et al., Plant Molec. Biol. 21: 673 (1993)(nucleotide sequence of the parsley ubi4-2 polyubiquitin gene).

J. Signal Transduction

Molecules stimulating signal transduction. See, e.g., Botella et al.,Plant Molec. Biol. 24: 757 (1994) (nucleotide sequences for mung beancalmodulin cDNA clones); and Griess et al., Plant Physiol. 104: 1467(1994) (nucleotide sequence of maize calmodulin cDNA clone).

K. Hydrophobic Moment Peptides

Hydrophobic moment peptides. See, e.g., PCT Application No. WO95/16776(peptide derivatives of Tachyplesin-inhibiting fungal plant pathogens)and PCT Application No. WO95/18855 (synthetic antimicrobial peptidesconferring disease resistance).

L. Membrane Permeases, Channel Formers, or Channel Blockers.

See, e.g., Jaynes et al., Plant Sci. 89: 43 (1993) (heterologousexpression of cecropin-β lytic peptide analog rendering transgenictobacco plants resistant to Pseudomonas solanacearum).

M. Viral-Invasive Proteins

Viral-invasive proteins or complex toxins derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparting resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as to related viruses. See, e.g., Beachy et al., Ann. Rev.Phytopathol. 28: 451 (1990). Coat protein-mediated resistance has beenconferred on transformed plants against alfalfa mosaic virus, cucumbermosaic virus, tobacco streak virus, potato virus X, potato virus Y,tobacco etch virus, tobacco rattle virus and tobacco mosaic virus. Id.

N. Insect-Specific Antibodies or Immunotoxins Derived Therefrom.

An antibody targeted to a critical metabolic function in the insect gutinactivating an affected enzyme, thereby killing the insect. Cf. Tayloret al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via single-chain antibody fragmentproduction).

O. Virus-Specific Antibodies

Virus-specific antibodies. See, e.g., Tavladoraki et al., Nature 366:469 (1993), (transgenic plants expressing recombinant antibody genes areprotected from virus attack).

P. Developmental-Arrestive Proteins

i. Developmental-Arrestive Proteins Produced by Pathogens or Parasites.See, e.g., Lamb et al., Bio/Technology 10: 1436 (1992) (fungal endoα-1,4-D-polygalacturonases facilitating fungal colonization and plantnutrient release by solubilizing plant cell wallhomo-α-1,4-D-galacturonase); and Toubart et al., Plant J. 2: 367 (1992)(cloning and characterization of a gene encoding beanendopolygalacturonase-inhibiting protein).

ii. Developmental-Arrestive Proteins Produced by Plants. See, e.g.,Logemann et al., Bio/Technology 10: 305 (1992) (increased resistance tofungal disease in transgenic plants expressing barleyribosome-inactivating gene).

Q. Genes Conferring Resistance to Herbicides

Herbicides inhibiting growing points or meristems, such as imidazolinoneor a sulfonylurea. Exemplary genes in this category encode mutant ALSand AHAS enzymes, respectively described by Lee et al., EMBO J. 7: 1241(1988); and Miki et al., Theor. Appl. Genet. 80: 449 (1990).

Glyphosate resistance (imparted by mutant5-enolpyruvl-3-phosphoshikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin acetyl transferase (bar) genes), andpyridinoxy or phenoxy proprionic acids and cyclohexones (ACCaseinhibitor-encoding genes). See, e.g., U.S. Pat. No. 4,940,835 to Shah etal., (EPSP clone conferring glyphosate resistance). A DNA moleculeencoding a mutant aroA gene can be obtained under ATCC Accession No.39256. The nucleotide sequence of such a mutant gene is disclosed inU.S. Pat. No. 4,769,061 to Comai. European Patent Application No. 0 333033 to Kumada et al. and U.S. Pat. No. 4,975,374 to Goodman et al.disclose nucleotide sequences of glutamine synthetase genes conferringresistance to herbicides such as L-phosphinothricin. A nucleotidesequence of a phosphinothricin-acetyl-transferase gene is disclosed inEuropean Patent Application 0 242 246 to Leemans et al. De Greef et al.,Bio/Technology 7: 61 (1989), describe the production of transgenicplants expressing chimeric bar genes coding for phosphinothricin acetyltransferase activity. Exemplary genes conferring resistance to phenoxyproprionic acids and cyclohexones, such as sethoxydim and haloxyfop, arethe Acc1-S1, Acc1-S2 and Acc1-S3 genes described by Marshall et al.,Theor. Appl. Genet. 83: 435 (1992).

Photosynthesis-inhibiting herbicides, such as triazines (psbA andgs+genes) and benzonitriles (nitrilase gene). Przibilla et al., PlantCell 3: 169 (1991) (transformation of Chlamydomonas using plasmidsencoding mutant psbA genes); U.S. Pat. No. 4,810,648 to Stalker(nucleotide sequences for nitrilase genes, available under ATCCAccession Nos. 53435, 67441 and 67442); Hayes et al., Biochem. J.285:173 (1992) (cloning and expression of DNA coding for glutathioneS-transferase).

R. Genes Conferring, or Contributing to, Value-Added Traits in Maize

i. Modified Fatty Acid Metabolism,

Fatty acid content can be modified by, for example, transforming a plantwith an antisense gene of stearoyl-ACP desaturase to increase stearicacid content. See, e.g., Knultzon et al., Proc. Natl. Acad. Sci. USA 89:2624 (1992).

ii. Decreased Phytate Content

Phytase-encoding genes enhancing breakdown of phytate by adding freephosphate to the transformed plant. See, e.g., Van Hartingsveldt et al.,Gene 127: 87 (1993) (nucleotide sequence of an Aspergillus niger phytasegene).

S. Genes Reducing Phytate Content.

For example, cloning, then reintroducing DNA associated with the alleleresponsible for maize mutants characterized by low levels of phyticacid. See, e.g., Raboy et al., Maydica 35: 383 (1990).

T. Modified Carbohydrate Compositions.

For example, transforming plants with a gene encoding an enzyme alteringstarch branching patterns. See, e.g., Shiroza et al., J. Bacteriol. 170:810 (1988) (nucleotide sequence of Streptococcus mutansfructosyltransferase gene); Steinmetz et al., Mol. Gen. Genet. 200: 220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene); Penet al., Bio/Technology 10: 292 (1992) (production of transgenic plantsexpressing Bacillus licheniformis α-amylase); Elliot et al., PlantMolec. Biol. 21: 515 (1993) (nucleotide sequences of tomato invertasegenes); Søgaard et al., J. Biol. Chem. 268: 22480 (1993) (site-directedmutagenesis of barley α-amylase gene); and Fisher et al., Plant Physiol.102: 1045 (1993) (maize endosperm starch branching enzyme II).

12. MAIZE TRANSFORMATION METHODS

Plant transformation methods contemplated to transform the inbred ofthis invention include biological and physical plant transformationprotocols. See, e.g., Miki et al., “Procedures for Introducing ForeignDNA into Plants” IN Methods in Plant Molecular Biology andBiotechnology, B. R. Glick and J. E. Thompson, Eds. (CRC Press, Inc.,Boca Raton, 1993) pages 67-88; Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology(expression vectors and in vitro culture methods for plant cell ortissue transformation and regeneration of plants); and B. R. Glick andJ. E. Thompson, Eds., CRC Press, Inc., Boca Raton, (1993) pages 89-119(expression vectors and in vitro culture methods for plant cell ortissue transformation and regeneration of plants).

A. Agrobacterium-Mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, e.g., Horsch etal., Science 227: 1229 (1985). A. tumefaciens and A. rhizogenes areplant pathogenic soil bacteria which infect, and genetically transform,plant cells during infection. The Ti and Ri plasmids of A. tumefaciensand A. rhizogenes, respectively, carry genes responsible for genetictransformation of the plant. See, e.g., Kado, Crit. Rev. Plant. Sci. 10:1 (1991). Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated gene transfer (transformation) are provided byGruber et al., “Vectors for Plant Transformation” IN Methods in PlantMolecular Biology and Biotechnology; Miki et al., “Procedures forIntroducing Foreign DNA into Plants” IN Methods in Plant MolecularBiology and Biotechnology, B. R. Glick and J. E. Thompson, Eds. (CRCPress, Inc., Boca Raton, 1993) pages 67-88; Moloney et al., Plant CellReports 8: 238 (1989); and U.S. Pat. No. 5,591,616 to Hiei et al.; andU.S. Pat. No. 6,822,144, issued 23 Nov. 2004 to Zhao et al.

B. Direct Gene Transfer

Despite the fact that the host range for Agrobacterium-mediatedtransformation is broad and with some exceptions in rice and maize, mostmajor cereal crop species and gymnosperms have generally beenrecalcitrant to this mode of gene transfer. Hiei et al., The PlantJournal 6: 271-282 (1994); and U.S. Pat. No. 5,591,616 to Hiei et al.Several methods of plant transformation, collectively referred to asdirect gene transfer, have been developed as alternatives toAgrobacterium-mediated transformation.

One generally applicable method of plant transformation ismicroprojectile-mediated transformation, wherein an expression vector isapplied to the surfaces of 1 to 41 m diameter microprojectiles. Theexpression vector is then introduced into plant tissues with a biolisticdevice which accelerates the microprojectiles to velocities sufficientto penetrate plant cell walls and membranes of the tissues, e.g., 300 to600 m/s. Sanford et al., Part. Sci. Technol. 5: 27 (1987); Sanford,Trends Biotech. 6: 299 (1988); Klein et al., Bio/Technology 6: 559-563(1988); Sanford, Physiol Plant 79: 206 (1990); Klein et al.,Biotechnology 10: 268 (1992); U.S. Pat. No. 5,550,318 to Adams et al.;U.S. Pat. No. 5,887,023 to Sautter et al; and U.S. Pat. Nos. 5,886,244and 5,990,387 to Tomes et al. In maize, several target tissues can bebombarded with DNA-coated microprojectiles to produce transgenic, hencederived, plants, including, for example, callus (Type I or Type II),immature embryos, and meristematic tissue.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9: 996 (1991). Alternatively,liposome or spheroplast fusion may be used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4: 2731 (1985); Christouet al., Proc Natl. Acad. Sci. U.S.A. 84: 3962 (1987), Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Hain et al., Mol. Gen. Genet.199: 161 (1985) and Draper et al., Plant Cell Physiol. 23: 451 (1982).Electroporation of protoplasts and whole cells and tissues has also beendescribed. Donn et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990); D'Halluin etal., Plant Cell 4: 1495-1505 (1992); Spencer et al., Plant Mol. Biol.24: 51-61 (1994); and U.S. Pat. No. 5,384,263 to Krzyzek et al.,previously referenced.

Following transformation of maize target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods known to the art.

The foregoing transformation methods may be used to produce transgenicderived inbred lines of this invention. These transgenic inbred linesmay then be crossed with another (non-transformed or transformed) inbredline to produce a transgenic hybrid maize plant. Alternatively, agenetic trait introgressed into a maize line using the foregoingtransformation protocols may be transferred to another line usingtraditional backcrossing techniques known to the plant breeding art,e.g., backcrossing an engineered trait from a public, non-elite lineinto an elite line, or from a hybrid maize plant with a foreigntransformed gene into an inbred line not containing that gene. As usedherein, “crossing” can refer to a single cross or to the process ofbackcrossing.

13. INDUSTRIAL APPLICABILITY

Maize is used as human food, livestock feed, as raw materials inindustry, and as a source of pigments. The food uses of maize, inaddition to human consumption of maize kernels, include products of thedry-milling and wet-milling industries, as well as pigments forcolorants. The principal products of maize dry milling are grits, mealand flour. The maize wet-milling industry provides maize starch, maizesyrup, and dextrose for food use. Maize oil is recovered from maizegerm, which is a by-product of both the dry-milling and wet-millingindustries. By way of illustration, and not limitation, a cup(approximately 164 grams) of corn would be expected to have thefollowing nutritional characteristics of Table I.

TABLE I EXEMPLARY CONSTITUENTS PRESENT IN MAIZE Nutrient Amount NutrientAmount Nutrient Amount calories 177.12 niacin equiv 3.26 mg alanine 0.48g calories from fat 18.90 vitamin B6 0.10 mg arginine 0.22 g caloriesfrom saturated fat 2.90 vitamin C 10.16 mg aspartate 0.40 g protein 5.44g vitamin E alpha 0.14 mg cystine 0.04 g equiv carbohydrates 41.18 gvitamin E IU 0.22 IU glutamate 1.06 g carbohydrates 41.18 g vitamin E mg0.80 mg glycine 0.22 g soluble fiber 0.18 g folate 76.10 mcg histidine0.14 g insoluble fiber 4.42 g vitamin K 0.66 mcg isoleucine 0.22 gsugar - total 4.26 g pantothenic acid 1.44 mg leucine 0.58 gmonosaccharides 1.32 g calcium 3.28 mg lysine 0.22 g disaccharides 2.78g copper 0.08 mg methionine 0.12 g fat - total 2.10 g magnesium 52.48 mgproline 0.48 g saturated fat 0.32 g manganese 0.32 mg serine 0.26 gmonounsaturated fat 0.62 g phosphorus 168.92 mg threonine 0.22 gpolyunsaturated fat 0.98 g potassium 408.36 mg tryptophan 0.04 g water114.10 g selenium 1.32 mcg tyrosine 0.20 g ash 1.18 g sodium 27.88 mgvaline 0.30 g vitamin A IU 355.88 IU zinc 0.78 mg vitamin A RE 36.08 RE18:0 stearic 0.02 g A - carotenoid 36.08 RE 18:1 oleic 0.62 g A - betacarotene 144.98 mcg 18:2 linoleic 0.96 g thiamin - B1 0.36 mg 18:3linoleic 0.02 g riboflavin - B2 0.12 mg omega 3 fatty acids 0.02 gniacin - B3 2.64 mg omega 6 fatty acids 0.96 g

Maize, including both grain and non-grain portions of the plant, is alsoused extensively as livestock feed, primarily for beef cattle, dairycattle, swine, and poultry. As will be shown, the grain and non-grainportions of the inbreds of this invention can be used as sources ofpigments. These pigments can be used in, without limitation, foods,beverages, and cosmetics.

Industrial uses of maize include production of ethanol, maize starch inthe wet-milling industry, and maize flour in the dry-milling industry.The industrial applications of maize starch and flour are based onfunctional properties, such as viscosity, film formation, adhesiveproperties, and abilities to suspend particles. Maize starch and flourhave applications in paper and textile industries. Other industrial usesinclude adhesives, building materials, foundry binders, laundrystarches, explosives, oil-well muds, and mining applications.

Plant parts other than the grain of maize are also used in industry. Forexample, stalks and husks are made into paper and wallboard and cobs areused for fuel and in making charcoal.

Hence, the seed of inbred maize line FAR601 or FAR045, the plantproduced from the inbred seed, the hybrid maize plant produced from thecrossing of the inbred, hybrid seed, and various parts of the hybridmaize plant and transgenic versions of the foregoing, can be utilizedfor human food, livestock feed, as a raw material in industry, or as asource of extracted pigments.

As previously indicated, the present invention relates to inbred linesof corn, including importantly plant parts and tissue of these inbredlines of corn, seed for such inbred lines, the use of the inbreds toproduce hybrid corn plants, hybrid corn plants obtained using at leastone of the inbreds as a parent and parts and tissue of these hybrid cornplants.

As used herein, the term “inbred,” “inbred line” or “inbred lines” meansa group of plants from a common ancestry which are essentiallyhomozygous and which are true breeding, i.e., uniform and stable withrespect to all of their agronomically important characteristics. Inpreferred embodiments, the inbred lines have the characteristics of thelines designated as FAR601 or FAR045. Generally, the preferred seeds ofinbred corn lines in accordance with the invention, have thecharacteristics of the seeds of each of the designated lines.

Anthocyanins are plant-based polyphenolic pigments belonging to theclass of molecules termed flavonoids. Consequently, anthocyanins are avery diverse group of compounds with a basic structure: anthocyanidinglycosidically linked to carbohydrate moieties and/or acyl groups.Anthocyanins commonly found in corn may include Cyanidin 3-glucoside,Cyanidin 3-(6″-malonylglucoside), Cyanidin 3-(6″-ethylmalonylglucoside),Cyanidin 3-(6″-dimalonylglucoside) (and other derivatives); Pelargonidin3-glucoside, Pelargonidin 3-(6-malonylglucoside), Pelargonidin3-(6″-ethylmalonylglucoside) (and other derivatives); Peonidin3-glucoside, Peonidin 3-(6″-malonylglucoside), Peonidin3-(6″-ethylmalonylglucoside) (and other derivatives); Cyanidin3-galactoside (and derivatives); Cyanidin 3-rutinoside (andderivatives); and Petunidin 3-glucoside (and derivatives).

Anthocyanins are widely found in flowering plants, such as corn. Thesecompounds are typically water soluble and non-toxic, displaying a rangeof colors from orange, bright red/purple to blue. Anthocyanins are polarmolecules and, therefore, are more soluble in polar solvents. Solubilityalso depends on various conditions such as pH, temperature, type ofsolvent, and concentration. Extraction may use solvents such as water,methanol, ethanol, or mixtures thereof, optionally acidified with anacid (e.g., between about 0.001% and 0.01% HCl or citric acid).

Other useful compounds may be present in extracts from the inbreds ofthis invention and their hybrid, such as other flavonoids, phenolicacids, and carbohydrates. Exemplary flavonoids include (−)-Epicatechin,(−)-Epicatechin 3-gallate, (−)-Epigallocatechin, (−)-Epigallocatechin3-gallate, (+)-Catechin, (+)-Gallocatechin. Exemplary phenolic acidsinclude Ferulic acid and derivatives, Quercetin and derivatives,P-coumaric acid and derivatives, Protocatechuic acid and derivatives,Vanillic acid and derivatives, Hesperitin and derivatives,Hydroxycinnamic acid and derivatives, Gallic acid and derivatives. Anonlimiting recital of carbohydrates would include common sugars such asarabinose, rhamnose or galactose and/or with acylating acids.

14. DESCRIPTION OF MAIZE INBREDS FAR601 AND FAR045

Each of the inbred corn lines FAR601 and FAR045, as indicated, aresubstantially homozygous and can be reproduced by planting seed of theline, growing the resulting corn plants under self-pollination orsibbing with adequate isolation, and harvesting the resulting seed usingtechniques familiar to those of skill in the art. A hybrid of FAR601 andFAR 045 can be produced by growing the two inbreds in proximity, thendetasseling one of the inbreds in this invention. Alternately, othermethods of pollen control as more fully described herein can be used aswell.

Each of the inbreds and their hybrid has shown uniformity and stabilitywithin the limits of environmental influence for each of the traits.Each of the instant inbreds and a three-way hybrid having the instantinbreds as two of its parents are described in Table IIA. A furtherdescription of the instant inbreds and their single cross hybrid aredescribed in Table IIB. As also indicated below, each inbred has beeneither sib-pollinated or self-pollinated a sufficient number generationswith careful attention paid to uniformity of plant type to ensurehomozygosity and phenotypic stability.

TABLE IIA MORPHOLOGICAL DESCRIPTION OF INBREDS FAR601 AND FAR045 AND ATHREE-WAY F1 HYBRID THEREOF Single Cross Male Female Female UPOVCharacteristic FAR601 FAR045 FAR044*FAR045 1. first leaf: anthocyanincoloration of sheet 9 9 9 2. first leaf: shape of tip 5 4 4 3. leaf:angle between blade and stem 5 2 3 4. attitude of blade 5 3 4 5. degreeof zig-zag 1 1 1 6. stem: anthocyanin coloration at base of 9 9 9 braceroots 7. time of anthesis 4 4 4 8. tassel: anthocyanin coloration atbase of 9 9 9 glume 9. tassel: anthocyanin coloration of glumes 8 5 710. tassel: anthocyanin coloration of anthers 5 1 4 11. tassel: densityof spikelets 3 3 3 12. tassel: angle between main axis and lateral 7 5 5branches 13. tassel: attitude of lateral branches 3 5 5 14. tassel:number of lateral branches 3 3 3 15. ear: time of silk emergence 4 4 416. ear: anthocyanin coloration of silks 9 3 9 17. ear: intensity ofanthocyanin coloration of 4 1 3 silks 18. leaf: anthocyanin colorationof sheath 9 1 9 19. tassel: length of main axis above lowest 6 6 6 sidebranches 20. tassel: length of main axis above highest 6 6 6 sidebranches 21. tassel: length of side branches 5 5 5 22. plant: length 6 67 23. ear: height of insertion, relative to plant 5 4 5 height 24. leaf:width of blade 4 5 5 25. ear: length of peduncle 3 2 2 26. ear: length 45 6 27. ear: diameter of ear 4 4 5 28. ear: shape 2 2 2 29. ear: numberof rows of grain 4 6 5 30. ear: type of grain 2 4 4 31. ear: color oftop of grain 9 9 9 32. ear: color of dorsal side of grain 9 9 9 33. ear:anthocyanin coloration of glumes of 9 9 9 cob 34. ear: intensity ofanthocyanin coloration of 9 5 7 glumes of cob

TABLE IIB MORPHOLOGICAL DESCRIPTION OF INBREDS FAR601 AND FAR045 ANDTHEIR SINGLE CROSS F1 HYBRID THEREOF UPOV Characteristic FAR045 FAR601FAR045*FAR601 1 first leaf: anthocyanin coloration of sheat 9 9 9 2first leaf: shape of tip 4 5 4 3 leaf: angle between blade and steam 2 53 4 attitude of blade 3 5 3 5 degree of zig-zag 1 1 1 6 stem:anthocyanin coloration at base of brace 9 9 9 roots 7 time of anthesis 44 4 8 tassel: anthocyanin coloration at base of glume 9 9 9 9 tassel:anthocyanin coloration of glumes 5 9 7 10 tassel: anthocyanin colorationof anthers 1 7 5 11 tassel: density of spikelets 3 3 3 12 tassel: anglebetween main axis and lateral 5 7 6 branches 13 tassel: attitude oflateral branches 5 3 5 14 tassel: number of lateral branches 3 3 5 15ear: time of silk emergence 4 4 4 16 ear: anthocyanin coloration ofsilks 1 9 9 17 ear: intensity of anthocyanin coloration of no 5 5 silks18 leaf: anthocyanin coloration of sheath 2 9 7 19 tassel: length ofmain axis above lowest side 6 6 7 branch 20 tassel: length of main axisabove highest side 6 6 7 branch 21 tassel: length of side branches 6 6 722 plant: length 6 6 7 23 ear: height of insertion, relative to plantheight 4 5 5 24 leaf: width of blade 5 4 6 25 ear: length of peduncle 33 3 26 ear: length 5 4 6 27 ear: diameter of ear 4 4 5 28 ear: shape 2 22 29 ear: number of rows of grain 6 4 6 30 ear: type of grain 4 3 4 31ear: color of top of grain 8 9 9 32 ear: color of dorsal side of grain 79 9 33 ear: anthocyanin coloration of glumes of cob 9 9 9 34 ear:intensity of anthocyanin coloration of 5 9 8 glumes of cob

The data present in Tables IIA and IIB represent 34 morphological traitsassociated with the Table of Characteristics published by theInternational Union for the Protection of New Varieties of Plants(UPOV). The data for Table IIA were gathered in Portugal during the 2006growing season. The data for Table IIB were also gathered in Portugal,but during the 2007 growing season. In each case, the data representratings of a person of skill in the art without resorting to thereference inbreds specified in the UPOV publication Guidelines for theConduct of Tests for Distinctness, Uniformity and Stability, TG/2/6,UPOV (1999). A more detailed description of these traits can be found inGuidelines for the Conduct of Tests for Distinctness, Uniformity andStability, TG/2/6, UPOV (1999).

As can be seen in the data of Tables IIA and IIB, each of the inbredvarieties and hybrids therefrom exhibits characteristic unusual strongred coloration in plant parts and tissue, including stems, leaves,glumes of cob and base of brace roots.

Tables IIIA and IIIB show the history of development for each of theinbred lines, each of which represents at least six to seven inbreedinggenerations.

TABLE IIIA INBRED LINE DEVELOPMENT AND GENEOLOGY OF FAR601 Line:FAR601 - Breeding History Selfing Cycle Country** Plot stage Activity2006/7 AR 06AC691049+50 S7 to S8 LM* and breeder seed production 2006 PT06AC090155/01+2 S6 to S7 inbred fixation 2005/6 NZ 05NZ-302/n S5 to S6inbred fixation 2004/5 NZ 04AR-151-54 S4 to S5 inbred fixation 2004 FRIN04-132/n# S4# inbred fixation 2003/4 NZ 03NZ-14/3 S3 to S4 inbredfixation 2003/3 CL 02CL-FM-25/1 S2 to S3 inbred fixation 2002 HU02HU5723/2 S1 to S2 inbred fixation 2001/2 CL 01CL-FM14/1 S0 to S1inbred fixation 2001 DE Rhine valley spontaneous red corn segregate asS0 *LM Line Maintenance; **AR Argentina, PT Portugal, HU Hungary, CLChile, DE Germany, NZ New Zealand, FR France

TABLE IIIB INBRED LINE DEVELOPMENT AND GENEOLOGY OF FAR045 Line:FAR045 - Breeding History Cycle Country** Plot Selfing stage Activity2006/7 AR 06FM691002 S7 to S8 LM* and breeder seed production 2006 PT06FM09007 S6 to S7 inbred fixation 2005/6 NZ 05NZ-305-02 S5 to S6 inbredfixation 2004/5 NZ 04AR-165/1 S4 to S5 inbred fixation 2004 FRIN04-145/2 S4 to S5 inbred fixation 2002 HU 02HU-5188/3 S3 to S4 inbredfixation 2001 HU 01HU-4010/1 S2 to S3 inbred fixation 2000/1 NZ00NZ-06/1 S1 to S2 inbred fixation 2000 HU 00HUFM-5287/6 S0 to S1 inbredfixation 1999 DE Rhine valley spontaneous red corn cross into elitegermplasm *LM Line Maintenance; **AR Argentina, PT Portugal, HU Hungary,CL Chile, DE Germany, NZ New Zealand, FR France

One aspect of the invention provides novel corn inbreds FAR601 andFAR045 as red-pigmented corn inbreds with superior characteristicsproviding excellent male and/or female parental lines for producing F₁corn hybrids also having excellent characteristics. It will beappreciated that the invention is intended to cover both inbred andhybrid plants and parts (tissues or cells) thereof. This includes anyplant parts and tissues acceptable for use in extracting pigmentspresent therein as well as for other uses described herein.

As seen in Table IIIA, line FAR601 was derived from Rhine Valleyspontaneous red corn segregate commonly available in Germany and wasinbred using pedigree breeding for seven generations. Inbred FAR601 isuniform and stable and appears to be homozygous for all characteristics.

As seen in Table IIIB, line FAR045 was derived from a cross of aspontaneous red corn into elite germplasm, then developed byself-pollination of successive inbreeding generations, accompanied byselection in a pedigree breeding program over 8 generations. Line FAR045has been observed to be uniform and stable and appears to be homozygousfor all agronomic characteristics.

A. WORKING EXAMPLE 1

One acre of the hybrid of FAR045*FAR601 was grown near Lamberton, Minn.in 2006. The grain yield, adjusted to 15.5% moisture at harvest andexpressed at 56 pounds per bushel, was 120 bushels per acre.

B. WORKING EXAMPLE 2

The hybrid FAR045*FAR601 was grown near Lamberton, Minn. in 2005 and2006. Tissues from representative plants were sampled to determineanthocyanin concentrations extracted from each tissue. The plants wereharvested near the black layer stage of maturity (late silage stage) onSep. 30, 2005 and Oct. 1, 2006. In each year, samples from the tissues,termed grain, husk, cob, leaf, and stalk, were taken, chopped by handand air dried at about 50 degrees Centigrade. The samples were thenground in a Waring Blender, then further ground in a Retsch Mill. Theground samples were subsequently immersed in an extraction solution orsolvent (50:50 water:ethanol, by volume having 0.1M HCl) and stirred oragitated at about 37 degrees Centigrade. After about 1.5 hours ofagitation in the extraction solution, the resulting slurry was filteredthrough a nylon mesh screen, then through #4 filter paper and assayedfor proportion of anthocyanins present. The portions of hybrid plantswere then assayed for anthocyanin content, the results being shown inTable IV.

TABLE IV Anthocyanin Proportions of Plant Tissues for Hybrid FAR045 ×FAR601 Plant Tissue Trait Grain Husk Cob Leaf Stalk Total 2006 Totalmass 51%  8%   7% 9% 25% 100% Lab pigment 0.22%   4.01%   0.69% 0.06%  0.27%    5.2% (% of mass) Harvested 2.8 0.4 0.4 0.5 1.4 5.5 mass (tons)Lab pigment 12.3 34.1 5.6 0.6 7.6 59.9 (lbs) Lab pigment 21% 57%   9% 1%13% 100% (% of total) 2005 Total mass 54%  7%   7% 8% 24% 100% Labpigment 0.166%   3.210%   1.240%  0.100%    0.459%    5.2% (% of mass)Harvested 2.8 0.4 0.4 0.2 0.6 4.3 mass (tons) Lab pigment 9.4 23.8 8.90.4 5.1 47.7 (lbs) Lab pigment 20% 50%   19% 1% 11% 100% (% of total)

In 2006 a greater amount, and percentage, of pigment was present.However, in both years most of the anthocyanin extracted was from thehusk leaves, followed by that from the grain. Other significant amountsof anthocyanin were obtained from the cob and stalk tissues and verylittle from the leaves.

C. WORKING EXAMPLE 3

The inbreds FAR045 and FAR601 were assayed by BioDiagnostics, Inc.,River Falls, Wis., for isozyme genotype using methods known to a personof ordinary skill in the art, e.g., protocols described by Stuber et al,“Techniques and Scoring Procedures for Starch Gel Electrophoresis ofEnzymes from Maize (Zea mays L.),” Technical Bulletin #286, NorthCarolina Agricultural Research Service, North Carolina State University,Raleigh, N.C. (1988). The results of this assay are shown in Table V.

TABLE V ISOZYME GENOTYPES OF FAR045 AND FAR601 Variety ADH1 ACP1 AMP1AMP3 GLU1 IDH1 IDH2 MDH1 FAR045 4/4 2/2 4/4 4/4, 4/5, 5/5 7/7 4/4 6/61/1, 1/6, 6/6 FAR601 4/4 4/4 4/4 3/3 7/7 4/4 4/4 6/6 Variety MDH2 MDH3MDH4 MDH5 PDG1 PDG2 PGM1 PGM2 FAR045 3/3 16/16 12/12 12/12 3.8/3.8 5/59/9 4/4 FAR601 6/6 16/16 12/12 12/12 3.8/3.8 5/5 9/9 4/4

The inbreds FAR045 and FAR601 had differing alleles for ACP1, AMP3,IDH2, AMP3, and MDH2. FAR045 was segregating at AMP3 with two allelespresent showing the expected two homozygotes and the singleheterozygote. However, a person of ordinary skill in the art willreadily recognize that FAR045 could be selected for homozygousindividuals at the AMP3 locus to thereby identify individuals breedingtrue for either of the alleles found. A person of ordinary skill in theart will also recognize that other FAR045 and FAR601 plants may haveother isozyme genotypes and nonetheless be within the spirit and scopeof this invention.

D. WORKING EXAMPLE 4

The composition of the seeds (kernels) of the inbreds FAR601, FAR045,their hybrid and another hybrid having a different, single cross femaleinbred parent, FAR045*FAR044 was determined. In each case, the male(pollen) parent was FAR601. The results of this assay are shown in TableVI.

TABLE VI KERNEL COMPOSITIONS OF INBREDS FAR045 AND FAR601 AND THEHYBRIDS FAR045*FAR601 AND FAR045*FAR044)(FAR601 Composition - KernelsOven Oil (oil- Moisture ether (ground) extraction) Protein StarchFAR045*FAR601 9.81% 4.34% 10.13% 74.1% FAR045*FAR044)(FAR601 10.75%4.18% 10.06% 73.2% FAR601 12.93% 4.90% 11.08% 70.1% FAR045 10.98% 3.76%12.48% 67.2%

Kernels of the two hybrids and two inbreds were as expected for oil andstarch percentages, but were higher than expected for proteinpercentages, based on yellow dent corn grades encountered in the upperMidwest of the United States. See, e.g., Corn: Chemistry and Technology,Stanley A. Watson and Paul E. Ramstad Editors, American Association ofCereal Chemists, Inc., St. Paul, Minn. (1987), page 72, Table IV.

E. WORKING EXAMPLE 5

Grain from the inbreds FAR045 and FAR601 and from the hybridFAR045*FAR601 was extracted using water:aceticacid:acetonitrile:trifluoroacetic acid (V/V) at respective ratios848:100:50:2. The kernels were extracted for six hours at ambienttemperature, then stored at a temperature of about 35 degrees F. forabout three days, then filtered. The extract was then subjected to highperformance liquid chromatography to determine proportions ofanthocyanin constituents and compared to the results of Working Example6 below. Results of this assay are presented in Table VII.

TABLE VII ANTHOCYANIN COMPONENTS OF FAR045, FAR601, AND FAR045*FAR601 %Area (080429 % Area by MPA Brunswick peak Ret FAR045* Extraction)FAR045* peak Ret no. Anthocyanin Time FAR601 FAR601 FAR045 FAR601 no.Time 1 cyanidin 3-glucoside 11.0 35.4 29.06 9.51 25.7 1 10.7 2 Unknown14.8 1.12 0.99 3 Unknown 16.1 1.30 1.00 1.13 4 Unknown 16.7 2.29 1.03 5cyanidin 3-(6″- 17.5 5.98 6.27 5.96 5.2 2 17.7 malonoyl)galactoside 6peonidin 3-glucoside 19.9 4.17 4.03 3.4 3 20.7 7 cyanidin 3-(6″- 22.032.37 33.22 35.73 42.4 4 24.4 malonoyl)glucoside 8 cyanidin 3- 26.0 1.051.25 17.61 2.8 5 30.6 (malonoyl)(malonoyl)galactoside 9 pelargonidin 3-26.7 1.21 1.44 2.50 2.0 6 32.2 (malonoyl)glucoside 10 Unknown 27.4 2.171.29 11 cyanidin 3- 28.3 5.76 9.19 21.13 7.6 7 33.6(malonoyl)(malonoyl)glucoside 12 peonidin 3- 30.2 3.63 4.29 4.2 8 35.8(malonoyl)galactoside peonidin 3-(malonoyl)glucoside 3.1 9 36.8 13pelargonidin 3- 34.3 0.49 0.49 0.6 10 38.5 (malonoyl)(malonoyl)glucoside14 cyanidin 3- 35.8 0.60 1.22 0.73 1.1 11 39.0(malonoyl)(succinoyl)galactoside peonidin 3- 0.8 12 39.5(malonoyl)(malonoyl)glucoside cyanidin 3- 0.6 13 39.9(malonoyl)(succinoyl)glucoside peonidin 3-(succinoyl)glucoside 0.3 1440.6 Total 97.54 94.77 94.30 99.8 Total % Non Acylated 39.57 33.09 9.5129.10 Total % Acylated 60.43 66.91 90.49 70.90 Total Peaks 19 33 11 14Total Area (relative concentration of 11000 54750 1570 Anthocyanin)Injection Volume ul 20 20 100

These results confirm that FAR601 produces very high amounts ofanthocyanins which may be extracted using only water as a solvent. Whilehaving a darkly red pigment, which is almost as intense at that ofFAR601, FAR045 produces a lower amount of water-extractibleanthocyanins. By way of illustration and not limitation, it is presentlypostulated that acylated anthocyanins are less soluble in water and maybe more tightly bound to plant tissues or plant cell components.Consequently, the higher concentrations of acylated anthocyanins presentin FAR045 may result in the lower anthocyanin concentrations present inthe extracts from this inbred.

F. WORKING EXAMPLE 6

Kernels from hybrids FAR045*FAR601 and FAR045*FAR044)(FAR601 and thevariety Kculli were assayed for anthocyanin content and constituents byBrunswick Laboratories, Norton, Mass. The assay compositions wereextracted from corn (maize) grain samples harvested in 2006 and 2007 andfrom the cultivar Kculli. The protocol included a first extraction of 40pounds (dry weight) of whole kernel corn using 60 pounds of deionizedwater at a low shear mixing action at 60 degrees Celsius for 1.5 hours.No other ingredients were present in the extracting solution other thandeionized water. The solution was then filtered from the corn and setaside. Another 60 pounds of deionized water was then added to the cornat low shear mixing at 60 degrees Celsius for another 1.5 hours. Thesolution was then filtered from the corn and combined with the firstextraction. The combined extracting solutions were then passed through aseries of progressively smaller filters ending with a 1 micron filter,at which point very little additional solid material could be filtered.The solution was then tested for percent solids (typically 0.4-0.6percent) and anthocyanin activity (typically 90-110 milligrams perliter). The filtered solution was evaporated approximately 100 times toa concentration of 40-60 percent solids and stabilized. In the casewhere the yield was one pound (454 grams) solids colorant, 50.4 grams ofa 50 percent solution of citric acid in deionized water was added tostabilize the product. The amount added was 10 percent citric acidsolution to a total citric acid solution and colorant. Anthocyaninconstituents were then determined by high performance liquidchromatography and are presented in Table VIII. Total anthocyaninconcentrations in the extract are shown in Table IX.

TABLE VIII ANTHOCYANIN CONSTITUENTS OF FAR045*FAR601,FAR045*FAR044)(FAR601, AND THE CULTIVAR KCULLI % anthocyanin FAR045*Anthocyanin FAR045*FAR044)(FAR601 FAR601 Kculli* cyanidin 3-glucoside13.7 25.7 43.1 cyanidin 3-(6″-malonoyl)galactoside 7.2 5.2 1.7 peonidin3-glucoside 2.5 3.4 5.7 cyanidin 3-(6″-malonoyl)glucoside 38.1 42.4 31.6cyanidin 3- 4.3 2.8 1.4 (malonoyl)(malonoyl)galactoside pelargonidin3-(malonoyl)glucoside 4.0 2.0 2.7 cyanidin 3- 13.4 7.6 2.1(malonoyl)(malonoyl)glucoside peonidin 3-(malonoyl)galactoside 5.5 4.24.4 peonidin 3-(malonoyl)glucoside 3.5 3.1 4.3 pelargonidin 3- 2.0 0.60.4 (malonoyl)(malonoyl)glucoside cyanidin 3- 2.1 1.1 0.9(malonoyl)(succinoyl)galactoside peonidin 3- 2.0 0.8 0.5(malonoyl)(malonoyl)glucoside cyanidin 3- 1.2 0.6 0.7(malonoyl)(succinoyl)glucoside peonidin 3-(succinoyl)glucoside 0.5 0.30.6 Essential Living Foods, Santa Monica, CA 90401.

TABLE IX TOTAL ANTHOCYANIN CONCENTRATIONS OF FAR045*FAR044)(FAR601,FAR045*FAR601, AND KCULLI Corn Hybrid or Variety Total Anthocyanin(mg/g) FAR045*FAR044)(FAR601 25.4 FAR045*FAR601 34.9 KCulli 52.0

G. WORKING EXAMPLE 7

Ground isolated pericarp of the hybrid FAR045*FAR601 was extracted usingwater, heat and agitation, then filtered and concentrated approximately10 fold using roto-evaporation. The analysis is presented in Table X.

TABLE X FAR045*FAR601 PERICARP CONSTITUENTS Total Anthocyanins 0.5-4%  Ash 1-4% Fat 0.5-4%   Protein 0.5-4%   Carbohydrates 10-30% Moisture54-87% *Anthocyanin calculation based on cyanidin-3-glucoside.*Composition analysis performed by Food Safety Net Services in SanAntonio, TX

H. WORKING EXAMPLE 8

Anthocyanins were extracted from seeds of FAR045, FAR601, and the hybridFAR045*FAR601 by the extraction protocol of Working Example 6. Percentactivity in terms of absorbance as measured by a spectrophotometer at510 nm and as based on the dry weight of the corn kernels. The resultsof this assay are shown in Table XI.

TABLE XI PERCENT ACTIVITY OF EXTRACTS OF INBRED FAR601 AND HYBRIDFAR045*FAR601 Percent Activity % Activity FAR045*FAR601 0.0271% FAR6010.1110%The aqueous protocol failed to extract sufficient quantities ofanthocyanins from FAR045. Accordingly, the assay could not be conductedfor extracts of FAR045 and no results are shown for this inbred. Theseresults show that FAR601 has high amounts of aqueously extractibleanthocyanins, higher than the hybrid FAR045*FAR601.

15. DEPOSITS

Applicant has made a deposit of at least 2500 seeds of Inbreds FAR045and FAR601 with the American Type Culture Collection (ATCC), Manassas,Va. 20110 USA, ATCC Deposit Nos. XXXXX and XXXXX. The seeds depositedwith the ATCC on Jun. 19, 2008 were taken from the deposit maintained byRed Rock Genetics, LLC, 41295 County Road 54, Lamberton, Minn. 56152since prior to the filing date of this application.

Access to these deposits will be available during the pendency of theapplication to the Commissioner of Patents and Trademarks and personsdetermined by the Commissioner to be entitled thereto upon request. Uponallowance of any claims in the application, the Applicant will make thedeposit available to the public pursuant to 37 C.F.R. § 1.808. Thesedeposits of the Inbreds FAR045 and FAR601 will be maintained in the ATCCdepository, which is a public depository, for a period of 30 years, or 5years after the most recent request, or for the enforceable life of thepatent, whichever is longer, and will be replaced if either becomesnonviable during that period. Additionally, Applicant has satisfied allthe requirements of 37 C.F.R. §§ 1.801-1.809, including providing anindication of the viability of the samples upon deposit. Applicant hasno authority to waive any restrictions imposed by law on the transfer ofbiological material or its transportation in commerce. Applicant doesnot waive any infringement of Applicant's rights granted under thispatent.

This invention has been described herein in considerable detail in orderto comply with the patent statutes and to provide those skilled in theart with the information needed to apply the novel principles and toconstruct and use embodiments of the example as required. However, it isto be understood that the invention can be carried out by specificallydifferent devices and that various modifications can be accomplishedwithout departing from the scope of the invention itself.

1. A seed of a corn cultivar designated FAR045, representative seed ofsaid cultivar deposited under ATCC Accession Number XXXXX.
 2. A cornplant, or a regenerable part thereof, produced by growing the seed ofclaim
 1. 3. A tissue culture of regenerable cells produced from theplant of claim
 2. 4. A protoplast produced from the tissue culture ofclaim
 3. 5. The tissue culture of claim 3, wherein cells of the tissueculture are from a plant part selected from a leaf, a pollen grain, anembryo, a root, a root tip, an anther, a pistil, an inflorescence, aseed, or a stem.
 6. A corn plant regenerated from the tissue culture ofclaim 3, said plant having all the morphological and physiologicalcharacteristics of corn cultivar FAR045, representative seed of saidcultivar deposited under ATCC Accession Number XXXXX.
 7. A method forproducing a hybrid corn seed wherein the method comprises crossing theplant of claim 2 with a different corn plant and harvesting theresultant hybrid corn seed.
 8. A hybrid corn seed produced by the methodof claim
 7. 9. A hybrid corn plant produced by growing the hybrid seedof claim
 8. 10. A method of producing an herbicide resistant corn plantwherein the method comprises transforming the corn plant of claim 2 witha transgene that confers herbicide resistance.
 11. An herbicideresistant corn plant produced by the method of claim
 10. 12. The cornplant of claim 11, wherein the transgene confers resistance to anherbicide selected from the group consisting of imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 13. A method of producing an insect resistant corn plantwherein the method comprises transforming the corn plant of claim 2 witha transgene that confers insect resistance.
 14. An insect resistant cornplant produced by the method of claim
 13. 15. The corn plant of claim 14wherein the transgene encodes a Bacillus thuringiensis endotoxin.
 16. Amethod of producing a disease resistant corn plant wherein the methodcomprises transforming the corn plant of claim 2 with a transgene thatconfers disease resistance.
 17. A disease resistant corn plant producedby the method of claim
 16. 18. A method of introducing a desired traitinto corn cultivar FAR045 wherein the method comprises: crossing FAR045plants grown from FAR045 seed, representative seed of which has beendeposited under ATCC Accession Number XXXXX, with plants of another cornline that comprise a desired trait to produce progeny plants, whereinthe desired trait is selected from the group consisting of malesterility, herbicide resistance, insect resistance and diseaseresistance; selecting progeny plants that have the desired trait toproduce selected progeny plants; crossing the selected progeny plantswith the FAR045 plants to produce backcross progeny plants; selectingfor backcross progeny plants that have the desired trait and the otherphysiological and morphological characteristics of corn cultivar FAR045to produce selected backcross progeny plants; and repeating saidcrossing and selecting for backcross progeny plants steps at least threetimes in succession, or, when using molecular markers, repeating saidcrossing and selecting at least once and selfing said backcross progenyplants at least once, to produce respective selected fourth or higherbackcross progeny plants or second-selfed backcross progeny plants thatcomprise the desired trait and essentially all of the otherphysiological and morphological characteristics of corn cultivar FAR045.19. A plant produced by the method of claim 18, wherein the plant hasthe desired trait and essentially all of the other physiological andmorphological characteristics of corn cultivar FAR045 as determined atthe 5% significance level when grown in the same environmentalconditions.
 20. The plant of claim 19 wherein the desired trait isherbicide resistance and the resistance is conferred to an herbicideselected from the group consisting of imidazolinone, sulfonylurea,glyphosate, glufosinate, L-phosphinothricin, triazine and benzonitrile.21. The plant of claim 19 wherein the desired trait is insect resistanceand the insect resistance is conferred by a transgene encoding at leasta portion of a Bacillus thuringiensis endotoxin.
 22. A process ofdeveloping a corn variety, comprising sequentially inbreedingsegregating generations of a corn hybrid having the plant of claim 2 asa parent until an advanced generation is attained, said advancedgeneration being F5 or greater.
 23. The process of claim 22, in whichinbreeding includes self-pollination.
 24. A process of extractinganthocyanin, comprising immersing a plant part from the plant of claim 2in a solvent to thereby extract anthocyanin from said plant part. 25.The process of claim 24, wherein said solvent is aqueous.
 26. Theprocess of claim 24, further comprising agitating said immersed plantpart.